Project Summary Antiretroviral treatment (ART) requires close monitoring of HIV-1 plasma viremia to confirm viral suppression, and to identify viral rebound due to nonadherence or drug resistance to initiate timely treatment modification. Thus, frequent efficient monitoring for HIV in blood via methods that are accessible for home self-testing is a critical need. Current self-detection strategies only detect antibody, and existing virus detection methods require extensive equipment and expertise (RNA) and/or have limited sensitivity (antigen). We aim to fill this gap with the development of an inexpensive, automated, portable system that will detect new or recurrent plasma viremia with high specificity and sufficient sensitivity for self-monitoring in these contexts. The ultimate goal of this project is to develop a platform technology for low-cost (< $2/test) point-of-care nucleic acid diagnostics for use on crude human samples (plasma) to quantitate viral RNA with a detection limit < 10 virus / sample (equivalent to a clinically-relevant threshold of 1000 copy/ml on 10 ul finger prick blood). The method will be simple to perform and will not require complex amplification procedures such as PCR. The novelty of our approach is to combine two innovative technologies invented by our team - one for target separation and concentration suitable for any starting blood volume, and one for label-free nucleic acid detection without amplification ? into an integrated device. The goal of our R66 is to design a laboratory-based integrated system to demonstrate quantitation of HIV viral load on tissue culture derived virus spiked into healthy plasma. The goals of our R33 are advanced development of our technology into a fully-automated handheld mobile-phone based device to be used by non-microfluidic experts, and evaluation using banked clinical samples through partnership with Penn's Center for AIDS Research.